β-Lactam antibiotics and their use

ABSTRACT

New β-lactam antibiotics of the formula ##STR1## in which A is hydrogen or methoxy, 
     B is phenyl optionally substituted by hydroxyl, halogen, methoxy, --CN and/or CH 3  --SO 2  --; thienyl; cyclohexenyl; 1,4-cyclohexadien-1-yl; or furyl, 
     E is oxygen or sulphur, 
     n is an integer having a value of 1 or 2, 
     Y is a group of the formula ##STR2## T is hydrogen, alkyl--CO--O--, hydroxyl, pyridinium, aminopyridinium, carbamoyloxy, azido, cyano, optionally N-substituted thiocarbamoylthio, optionally substituted-S-phenyl, or a --S--Het group in which Het represents an optionally substituted heterocyclic 5-membered or 6-membered ring, 
     U is oxygen, sulphur or --CH 2  --, 
     Z is a group of the formula ##STR3## R 1 , R 2  and R 3  each is hydrogen or various organic radicals, i.e. compounds wherein Z is an olefinic or hydrated olefinic radical directly connected to the ring nitrogen, or salts thereof, are antibacterially active and useful as antibiotics in pharmacy, as animal feed supplements for promoting growth, and as preservatives.

The present invention relates to certain new β-lactam compounds, to aprocess for their production and to their use as medicaments, inparticular as antibacterial agents and as agents for promoting growthand for improving feedstuff utilization in animals.

β-lactam compounds substituted in a particular manner, such asα-(imidazolidin-2-oxo-1-yl-carbonylamino)-benzylpenicillins substitutedin a particular manner, and corresponding cephalosporins having animidazolidin-2-oxo-1-yl-carbonylamino side chain are known from GermanOffenlegungsschriften (German Published Specifications) Nos. 2,104,580,2,152,967 2,258,973, 2,402,465 and 2,428,139.

The present invention provides compounds which are β-lactams of thefollowing general formula I or their salts: ##STR4## in which A ishydrogen or methoxy;

B is phenyl optionally substituted by hydroxyl, halogen, methoxy, --CNand/or CH₃ --SO₂ --; thienyl; cyclohexenyl; 1,4-cyclohexadien-1-yl; orfuryl;

E is oxygen or sulphur;

n is an integer having a value of 1 or 2;

Y is a group of the formula ##STR5## in which the carbon atom whichcarries the carboxyl group is bonded to the nitrogen atom of theβ-lactam ring and T is hydrogen, alkyl--CO--O--, hydroxyl, pyridinium,aminopyridinium, carbamoyloxy, azido, cyano, thiocarbamoylthio,optionally substituted --S--phenyl or a --S--Het group, in which Hetrepresents an optionally substituted heterocyclic 5-membered or6-membered ring, and

U is oxygen, sulphur or --CH₂ --; and

Z is a group of the formula ##STR6## wherein R₁, R₂ and R₃ are the sameor different and each is hydrogen or alkyl having from 1 to 5 C atomsand being optionally substituted, or

R₁ and R₂ or R₂ and R₃, together with the C atom to which they arebonded, represent an optionally substituted ring having from 2 to 7 Catoms and optionally containing, as ring members, 1 or 2 groups, whichare the same or different, and each of which is --O--, --S--, --CO--,--SO--, --SO₂ -- or --N--R' wherein

R' is hydrogen, lower alkyl or lower alkylcarbonyl, said β-lactams ofthe formula I being in either of the two possible configurations R and Sin respect of the chirality center C* or in the form of mixtures in saidR and S configurations, and optionally in the form of mixtures of thediastereomers resulting therefrom.

The new β-lactam compounds according to the invention differ from theknown compounds of the state of the art in that for example, in theimidazolidinone radical the 3-position N is directly bonded, that is tosay without an intermediate member, such as, for example, a carbonyl orsulphonyl group, to an olefinic C atom or a hydrate thereof.

The compounds according to the invention have powerful antibacterialproperties and possess the property of improving growth and feedstuffutilization in animals.

In a further aspect the present invention provides a process for theproduction of a compound of the invention in which a compound of theformula II: ##STR7## wherein A, B C* and Y have the same meaning asdefined hereinbefore in formula I; or a derivative thereof obtained bysilylation on the carboxyl group or on the carboxyl and primary aminogroup, is reacted with a compound of the general formula III: ##STR8##in which Z, E and n have the same meaning as defined hereinbefore and

W is halogen, azide or another nucleofugic leaving group,

optionally in the presence of a solvent and/or as acid-binding agent,preferably at a temperature of from -20° C. to +50° C., and, where asilylated derivative of a compound of formula II has been used,splitting off the silyl group(s) and optionally converting the resultingβ-lactam of formula I or salt thereof into a salt thereof or thecorresponding free β-lactam of formula I, respectively.

Surprisingly, the new β-lactam compounds exhibit a very good activityagainst a broad spectrum of pathogens, in addition to being welltolerated. The substances according to the invention thus represent anenrichment of pharmacy.

If D-α-amino-benzylpenicillin and1-chlorocarbonyl-2-oxo-3-propenyl-imidazolidine are used as startingmaterials, the course of the reaction can be represented by thefollowing reaction equation: ##STR9##

In the formulae I and III, the unsubstituted radicals Z preferablycontain from 2 to 7 carbon atoms. Examples which may be mentioned are:vinyl, propenyl, butenyl, isobutenyl, sec.-butenyl, 1-methylisobutenyland the cyclic radicals: ##STR10## and their hydrated forms.

Z can be monosubstituted, disubstituted or trisubstituted, preferablymonosubstituted or disubstituted, in particular monosubstituted.Examples of substituents which may be mentioned are: lower alkyl, inparticular methyl, ethyl, propyl, isopropyl and t-butyl, preferablymethyl; lower alkylidene, in particular methylene, ethylidene andisopropylidene; vinyl, propenyl allyl and isopropenyl; loweralkoxymethyl, in particular methoxymethyl; lower alkylthiomethyl, inparticular methylthiomethyl; trifluoromethyl; hydroxymethyl; formyl;lower alkanoyl, in particular acetyl; lower alkanoyloxymethyl, inparticular acetoxymethyl; benzyl; aryl, in particular phenyl;cyanomethyl; the CH₃ --NH--CO--CH₂ -- and (CH₃)₂ N--CO--CH₂ -- groups;lower alkoxycarbonyl, in particular methoxycarbonyl and ethoxycarbonyl;carboxyl; cyano; hydroxyl; lower alkanoyloxy, in particular acetoxy;lower alkoxy, in particular methoxy and ethoxy; benzyloxy; halogen, inparticular fluorine, chlorine and bromine, preferably chlorine;mercapto; lower alkylthio, in particular methylthio and ethylthio; loweralkylsulphinyl, in particular methylsulphinyl and ethylsulphinyl; loweralkylsulphonyl, in particular methylsulphonyl and ethylsulphonyl; andthe CH₃ --CO--NH--, CH₃ --CO--N--(CH₃)--, CH₃ --SO₂ --NH-- and ##STR11##groups.

The alkenyl radicals Z are preferably unsubstituted.

A particularly preferably represents hydrogen and E particularlypreferably represents oxygen. U particularly preferably denotes sulphur.

In the definition of T, alkyl in alkyl--CO--O-- preferably denotes alkylwith 1 to 4, in particular 1 or 2, carbon atoms. Examples which may bementioned are methyl and ethyl, preferably methyl. T particularlypreferably represents hydrogen, OH or methyl--CO--O--.

The thiocarbamoylthio radical (in the definition of T) can besubstituted on the N atom by one or two lower alkyl radicals.Furthermore, this N atom can be a constituent of a pyrrolidine,piperidine, morpholine and N⁴ lower alkylpiperazine ring.

The heterocyclic ring Het in --S--Het (in the definition of T) consistsof 5 or 6 ring members and contains from 1 to 4, preferably from 2 to 4,and very particularly preferably 3 or 4, identical or differenthetero-atoms, each of which is oxygen, sulphur or nitrogen. Theheterocyclic ring is preferably unsaturated and particularly preferablycontains 2 double bonds. The heterocyclic ring can contain one or more,preferably 1 or 2, in particular one, substituents. Examples ofsubstituents which may be mentioned are: halogen, such as fluorine,chlorine, bromine and iodine, preferably chlorine or bromine; amino,lower alkylamino, and di-lower alkylamino; lower alkyl; cycloalkyl(having from 3 to 7 preferably 5 or 6 carbon atoms in the cycloalkylmoiety), lower alkyloxy; trifluoromethyl; phenyl; benzyl; and acylaminowith preferably 2 to 5, in particular 2 or 3, carbon atoms. Particularlypreferred examples of --S--Het which may be mentioned are: ##STR12## inwhich R¹ is H, CH₃ or C₂ H₅ ; and X is O or S, ##STR13## in which R² isH, CH₃, CF₃, NHCH₃, NHCHO, N(CH₃)₂ or NHCOCH₃ ; and X is O or S; and##STR14## in which R³ is H or CH₃.

The --S-- phenyl radical in the definition of T can carry one or more,preferably from 1 to 3, in particular 1 or 2, identical or differentsubstituents, preferred substituents being those which are listed aboveas being possible substituents of the radical --S--Het.

Nucleofugic leaving groups in the definition of W are to be understoodas all the nucleofugic groups customarily used in organic chemistry, andabove all those which are described in Angewandte Chemie, 81 (1969),page 543.

The phenyl radical B can carry one or more identical or different,preferably from 1 to 3, in particular 1 or 2 and most preferably 1,substituent(s) each of which is hydroxyl, halogen, methoxy, --CN-- orCH₃ --SO₂ --. In the case of monosubstitution, the substituent ispreferably in the 4-position (relative to the bond of the phenyl radicalon the asymmetric carbon atom C*).

Halogen as a substituent in the phenyl radical B denotes fluorine,chlorine, bromine and iodine, preferably fluorine, chlorine or bromine,in particular fluorine or chlorine.

B particularly preferably represents phenyl, 4-hydroxyphenyl,1,4-cyclohexadien-1-yl and furyl. Among the new β-lactam salts of theinvention, those salts that are pharmaceutically acceptable areparticularly important and are preferred.

Pharmaceutically acceptable salts of the compounds of the formula I aresalts of these compounds with inorganic and organic bases on the acidcarboxyl group or on the acid carboxyl and sulphonyl groups. Bases whichcan be employed for this are all the bases customarily used inpharmaceutical chemistry, in particular in the chemistry of antibiotics.Examples of inorganic bases which may be mentioned are: alkali metalhydroxides and alkaline earth metal hydroxides, alkali metal carbonatesand alkaline earth metal carbonates and alkali metal bicarbonates, suchas sodium hydroxide and potassium hydroxide, calcium hydroxide andmagnesium hydroxide, sodium carbonate and potassium carbonate, calciumcarbonate, and sodium bicarbonate and potassium bicarbonate; andaluminum hydroxide and ammonium hydroxide. Organic bases which can beused are primary, secondary and tertiary aliphatic amines andheterocyclic amines. Examples which may be mentioned are: di- andtri-lower alkylamines, for example diethylamine and triethylamine,tri-β-hydroxyethylamine, procaine, dibenzylamine,N,N'-dibenzylethylenediamine, N-benzyl-β-phenyl-ethylamine, N-methyl-and N-ethyl-morpholine, 1-ephenamine, dehydroabietylamine,N,N'-bis-dehydroabietylethylenediamine and N-lower alkylpiperidine.So-called basic aminoacids, such as lysine or arginine, can also beadvantageously used as bases. Particularly preferred salts are thesodium salts.

By the expression "lower alkyl" there is to be understood herein, ineach case, straight-chain or branched alkyl having from 1 to 5,preferably 1 to 3, in particular 1 or 2 carbon atoms. In connection withother groups, such as in "di-lower alkylamino", the above definition of"-lower alkyl-" only relates to the alkyl moiety of the group concerned.

Particularly preferred compounds of the general formula I are those inwhich

A is hydrogen;

B is phenyl, hydroxyphenyl (particularly preferably p-hydroxyphenyl),1,4-cyclohexadien-1-yl or furyl;

E is oxygen;

Y is a group of the formula ##STR15## wherein T is --O--CO--CH₃, --OH,1-methyl-tetrazol-5-yl-thio, 2-methyl-1,3,4-thiadiazol-5-yl-thio,3-methyl-1,2,4-thiadiazol-5-yl-thio or2-trifluoromethyl-1,3,4-thiadiazol-5-yl-thio and

Z is vinyl, propenyl, isopropenyl, isobutenyl, sec.-butenyl or1-methylisobutenyl, or a hydrated form thereof, and

C* is in the D--═R-configuration,

and the salts, especially the sodium salts of these compounds.

All the crystal forms and hydrate forms of the compounds of the generalformula I according to the invention and their salts are antibacteriallyactive in the same manner.

The compounds of the general formula II which can be used according tothe invention are already known or are obtainable by known methods(compare, for example, E. H. Flynn, Cephalosporins and Penicillins,Academic Press, New York and London, 1972).

Examples which may be mentioned are: α-aminobenzyl-penicillin (shortname: ampicillin), α-amino-p-hydroxybenzyl-penicillin (short name:amoxicillin), α-amino-p-methylbenzyl-penicillin,α-amino-p-chlorobenzylpenicillin,6-(2-amino-2-(1,4-cyclohexadien-1-yl)-acetamido)-penicillanic acid(short name: epicillin),7-(α-amino-phenylacetamido)-3-methyl-ceph-3-em-4-carboxylic acid,7-(β-amino-phenylacetamido)-3-acetoxy-methyl-ceph-3-em-4-carboxylic acid(short name: cephaloglycine),7-(α-amino-phenylacetamido)-3-((1-methyltetrazol-5-yl)-thiomethyl)-ceph-3-em-4-carboxylicacid,7-(α-amino-phenylacetamido)-3-((2-methyl-1,3,4-thiadiazol-5-yl)-thiomethyl)-ceph-3-em-4-carboxylicacid as well as the sodium salts and the mono- and di-(trimethylsilyl)derivatives of these compounds.

All the crystal forms, hydrate forms and salts of the compounds of thegeneral formula II are suitable starting materials for the processaccording to the invention.

The compounds of the general formula III used as starting materials areknown or are obtainable by known methods. For example, they can beprepared in the customary manner by reacting a heterocyclic compound ofthe general formula IV: ##STR16## in which n is 1 or 2 and Z has thesame meaning as defined hereinbefore in formula I, with, for example, amolar amount of a compound of the general formula W--CE--W in which Wand E have the same meaning as defined hereinbefore in formula III, suchas, for example, phosgene or thiophosgene, in an inert organic solvent,such as, for example, tetrahydrofuran, or in a mixture of water and aninert organic solvent such as, for example, chloroform, at a temperatureof from 0° to 25° C. optionally in the presence of a molar amount of abase, such as, for example, triethylamine, and by customary working upand purification.

The preparation of the respective compounds of the general formula IVand III is indicated in each case in the examples, unless they aredescribed in the literature. In general the remaining starting compoundsare all readily available in an analogous manner.

Examples of compounds of formula III which may be mentioned are:

    ______________________________________                                         ##STR17##                                                                                     ##STR18##                                                    Z'            W'       Z'             W'                                      ______________________________________                                        Vinyl         Cl       Vinyl          Cl                                      Propenyl      Cl       Propenyl       Cl                                      Isobutenyl    Cl       Isobutenyl     Cl                                      sec.-Butenyl  Br       sec.-Butenyl   N.sub.3                                 1-Methyl-isobutenyl                                                                         N.sub.3  1-Methyl-isobutenyl                                                                          Br                                      ______________________________________                                    

Diluents which can be used in the reaction according to the invention,of the compounds of the formulae II and III are water and virtually allthe customary inert organic solvents, optionally mixed with water.

If the non-silylated compounds of the general formula II are used forthe reaction with the compounds of the general formula III, thisreaction can be carried out, for example, in any desired mixtures ofwater and those organic solvents which are water-miscible, for example,ketones, such as acetone, cyclic ethers, for example tetrahydrofuran anddioxane, nitriles, for example acetonitrile, formamides, for exampledimethylformamide, dimethylsulphoxide or alcohols, for example,isopropanol. In this procedure, the pH of the reaction mixture is kept,for example, at from 6.5 to 8.0 by adding bases or using buffersolutions (for example phosphate buffers or citrate buffers). However,the process according to the invention can also be carried out at a pHof from pH 1.5 to pH 9.5, for example from 4.5 to 9.0, or at from pH 2.0to 4.5. Furthermore, it is possible to carry out the reaction in asolvent which is not water-miscible, for example a halogenatedhydrocarbon, such as chloroform or methylene chloride, a base,preferably triethylamine, diethylamine, or N-ethylpiperidine, beingadded desirably. Furthermore, the reaction can be carried out in amixture of water and a solvent which is not water-miscible, such as, forexample an ether, for example diethyl ether, halogenated hydrocarbon,for example, chloroform and methylene chloride, carbon disulphide,ketone, for example isobutyl methyl ketone, ester, for example ethylacetate, aromatic solvent, for example benzene, and the like, it beingappropriate to stir the mixture vigorously and to keep the pH value atfrom 1.5 to 9.5, preferably from 4.5 to 9.0 or, for example, from 2.0 to3.0, by adding bases or using buffer solutions. However, it is alsopossible to carry out the reaction in water alone in the absence of anorganic solvent in the presence of an organic or inorganic base or withthe addition of buffer substances.

Suitable silyl radicals for the derivatives of the compounds of thegeneral formula II are all the silyl groups which are described in theliterature and are used for similar purposes. It is known of some ofthese silyl groups, for example, of the trimethylsilyl group, that theyare rapidly split off by water or solvents containing HO groups. On theother hand, it is known of other silyl groups, for example of thedimethoxy-methyl-silyl group or of the dimethyl-tert.-butyl-silyl group,that they are substantially more resistant towards hydrolysis. Dependingon the nature of the silyl radicals used, the reaction may thereforehave to be carried out in a solvent which is completely anhydrous andfree from hydroxyl groups. If monosilylated compounds of the generalformula II are used for the reaction with the compounds of the generalformula III, it is possible to carry out the reaction with or without,though preferably with, the addition of a base.

If disilylated derivatives of compounds of the general formula II areused, it is possible to carry out the reaction with or without, thoughpreferably without, the addition of a base.

All the inorganic and organic bases customarily employed in organicchemistry can be used as the base including bases such as alkali metalhydroxides and alkaline earth metal hydroxides, alkaline earth metaloxides, alkali metal carbonates and alkaline earth metal carbonates andbicarbonates, ammonia, primary, secondary and tertiary aliphatic andaromatic amines and heterocyclic bases. Examples which may be mentionedare sodium hydroxide, potassium hydroxide and calcium hydroxide, calciumoxide, sodium carbonate and potassium carbonate, sodium bicarbonate andpotassium bicarbonate, ethylamine, methyl-ethylamine, triethylamine,hydroxyethylamine, aniline, pyridine and piperidine.

Suitable buffer mixtures are all the buffer mixtures customarily used inorganic chemistry, such as phosphate buffers, citrate buffers andtris-(hydroxymethyl) aminomethane buffers.

In the reaction of the compounds of formulae II and III the reactiontemperatures can be varied within a relatively wide range.

In general, the reaction is carried out at from -20° C. to +50° C.,preferably from 0° to +25° C. As in the case of most chemical reactions,temperatures which are higher or lower than those indicated can also beused. However, if the values indicated are exceeded considerably, sidereactions which lower the yield or have an adverse influence on thepurity of the products take place to an increasing extent. On the otherhand, reaction temperatures which are lowered excessively reduce therate of reaction so severely that reductions in yield can occur.

The reaction can be carried out under normal pressure, but also underreduced or elevated pressure. In general, the reaction is carried outunder ambient pressure.

In carrying out the process according to the invention, the reactantscan be reacted with one another in equimolar amounts. However, incertain circumstances, it can be appropriate to use one of the tworeactants in excess, in order to facilitate the purification of thedesired β-lactam antibiotic or its preparation in the pure state, and toincrease the yield.

For example, it is possible to employ the reactants of the generalformula II in an excess of from 0.1 to 0.3 molar equivalents and therebyto achieve less decomposition of the reactants of the general formulaIII in a water-containing solvent mixture. Because of their goodsolubility in aqueous mineral acids, the excess of the reactants of thegeneral formula II can be easily removed during the working up of thereaction mixture.

On the other hand, however, the reactants of the general formula III canalso be advantageously employed in an excess of, for example, 0.1 to 1.0molar equivalents. By this procedure, the reactants of the generalformula II are better utilized and the decomposition of the reactants ofthe general formula III, which proceeds as a side reaction inwater-containing solvents, is compensated. Since the compounds of thegeneral formula III added in excess are rapidly converted in water intoneutral nitrogen-containing heterocyclic compounds, which can be easilyremoved, the purity of the β-lactam compounds is scarcely impaired bythis procedure.

Bases can be added in equimolar amounts, but also in excess, withrespect to the reactants of the formulae II and III. The amount of baseused depends, of course, on the chosen range of pH values to bemaintained.

The working up of the reaction mixtures for the preparation of theβ-lactam compounds according to the invention and their salts and thepurification of these compounds may be carried out in the manner whichis generally customary for penicillins and cephalosporins, for example,by removing the solvent, taking up the residue in an organic solvent(s)and precipitating and recrystallizing the products.

The sodium salts are obtained from an ethereal solution of the freeacids in a particularly advantageous manner by precipitation with sodium2-ethylhexanoate.

If the monosilylated or disilylated compounds of the general formula IIare used in the reactions according to the invention, the hydrolyticsplitting off of the silyl radicals or radical occurs in the course ofthe aqueous working up of the reaction mixtures, optionally at an acidpH.

Specific new active compounds in accordance with the invention includethe following compounds of formulae V, VI, VII and VIII:

    __________________________________________________________________________     ##STR19##                            V                                       Z                    B                                                        __________________________________________________________________________    CH.sub.2CH           Phenyl                                                   "                    4-Hydroxyphenyl                                          CH.sub.3CHCH         Phenyl                                                   "                    4-Hydroxyphenyl                                          "                    4-Cyanophenyl                                             ##STR20##           Phenyl 4-Hydroxyphenyl                                    ##STR21##           Phenyl 4-Hydroxyphenyl                                   "                    4-Cyanophenyl                                            "                    Furfuryl                                                  ##STR22##           Phenyl                                                   "                    4-Hydroxyphenyl                                          __________________________________________________________________________     ##STR23##                            VI                                      Z             B        T                                                      __________________________________________________________________________    CH.sub.2CH    Phenyl   OCOCH.sub.3                                            "             4-Hydroxyphenyl                                                                        "                                                      "             4-Cyanophenyl                                                                          "                                                      "             Furfuryl "                                                       ##STR24##    Phenyl   "                                                      "             4-Hydroxyphenyl                                                                        "                                                      CH.sub.3CHCH  Phenyl   OCOCH.sub.3                                            "             "                                                                                       ##STR25##                                             "             "                                                                                       ##STR26##                                             "             4-Hydroxyphenyl                                                                        OCOCH.sub.3                                            "             "                                                                                       ##STR27##                                             "             4-Hydroxyphenyl                                                                         ##STR28##                                             "             4-Cyanophenyl                                                                          OCOCH.sub.3                                            "             "                                                                                       ##STR29##                                             "             "                                                                                       ##STR30##                                             "             Furfuryl OCOCH.sub.3                                            "             "                                                                                       ##STR31##                                              ##STR32##    Phenyl                                                          "             "        OCOCH.sub.3                                            "             "                                                                                       ##STR33##                                             "             "                                                                                       ##STR34##                                             "             Hydroxyphenyl                                                                          OCOCH.sub.3                                            "             "                                                                                       ##STR35##                                             "             "                                                                                       ##STR36##                                              ##STR37##    Furfuryl OCOCH.sub.3                                                                    ##STR38##                                             __________________________________________________________________________     ##STR39##                            VIII                                    Z             B        T                                                      __________________________________________________________________________    CH.sub.3CHCH  Phenyl   OCOCH.sub. 3                                            ##STR40##    "        "                                                       ##STR41##    "        "                                                      __________________________________________________________________________     ##STR42##                                                                    Z                    B                                                        __________________________________________________________________________    CH.sub.2CH           Phenyl                                                   CH.sub.3CHCH         Phenyl                                                   "                    p-Hydroxyphenyl                                          __________________________________________________________________________    Z                    B                                                        __________________________________________________________________________     ##STR43##           Phenyl                                                   "                    p-Hydroxyphenyl                                           ##STR44##           Phenyl                                                   __________________________________________________________________________     ##STR45##                                                                    Z                    B                                                        __________________________________________________________________________    CH.sub.2CH           Phenyl                                                   CH.sub.3CHCH         Phenyl                                                   "                    p-Hydroxyphenyl                                           ##STR46##           Phenyl                                                   "                    p-Hydroxyphenyl                                           ##STR47##           Phenyl                                                   __________________________________________________________________________

If in the radicals Z in the compounds of general formulae I and III oneor more asymmetric centers or cis- and trans-forms occur, it should beunderstood that all the possible R-, S-, cis- and trans-forms and allthe possible combinations of these forms of the compounds of generalformulae I and III are included in references to compounds of saidformulae unless the contrary is stated.

The compounds according to the invention display a powerfulantimicrobial activity, coupled with low toxicity and good tolerance.These properties enable the compounds to be used as active ingredientsin medicine and as substances for preserving inorganic and organicmaterials, especially organic materials of all kinds, for examplepolymers, lubricants, paints, fibers, leather, paper and timber, and offoodstuffs and water.

The compounds according to the invention are active against a very broadspectrum of micro-organisms. With their aid it is possible to combat,for example, Gram-negative and Gram-positive bacteria and bacteria-likemicro-organisms and to prevent, alleviate and/or cure diseases caused bythese pathogens.

The compounds according to the invention are particularly active againstbacteria and bacteria-like micro-organisms. They are thereforeparticularly suitable for the prophylaxis and chemotherapy of local andsystemic infections caused by these pathogens, in human medicine andveterinary medicine.

For example, local and/or systemic diseases which are caused by thefollowing pathogens or by mixtures of the following pathogens can betreated and/or prevented:

Micrococcaceae, such as Staphylococci, for example Staphylococcusaureus, Staph. epidermidis, Staph. aerogenes and Gaffkya tetragena(Staph.=Staphylococcus);

Lactobacteriaceae, such as Streptococci, for example Streptococcuspyogenes, α- and β-haemolyzing Streptococci, non-(γ-)-haemolyzingStreptococci, Str. viridans, Str. faecalis (Enterococci), Str.agalactiae, Str. lactis, Str. equi. Str. anaerobis and Diplococcuspneumoniae (Pneumococci) (Str.=Streptococcus);

Neisseriaceae, such as Neisseriae, for example Neisseria gonorrhoeae(Gonococci), N. meningitidis (Meningococci), N. catarrhalis and N. flava(N.=Neisseria);

Corynebacteriaceae, such as Corynebacteria, for example Corynebacteriumdiphtheriae, C. pyogenes, C. diphtheroides and C. acnes(C.=Corynebacterium);

Enterobacteriaceae, such as Escherichiae bacteria of the Coli group,Escherichia bacteria, for example, Escherichia coli, Enterobacterbacteria, for example E. Aerogenes and E. cloacae, Klebsiella bacteria,for example K. pneumoniae and K. ozaenae, and Serratia, for exampleSerratia marcescens (E.=Enterobacter) (K.=Klebsiella), Proteae bacteriaof the Proteus group, Proteus, for example Proteus vulgaris, Pr.morganii, Pr. rettgeri and Pr. mirabilis (Pr.=Proteus), Providencia, forexample Providencia sp., Salmonelleae, Salmonella bacteria, for exampleSalmonella paratyphi A and B, S. typhi, S. enteritidis, S. cholerae suisand S. typhimurium (S.=Salmonella), and Shigella bacteria, for exampleShigella dysenteriae, Sh. ambigua, Sh. flexneri, Sh. boydii and Sh.sonnei (Sh.=Shigella); Pseudomonadaceae, such as Pseudomonas bacteria,for example Pseudomonas aeruginosa, and Aeromonas bacteria, for exampleAeromonas liquefaciens, Spirillaceae, such as Vibrio bacteria, forexample Vibri cholerae, Parvobacteriaceae or Brucellaceae, such asPasteurella bacteria, for example Pasteurella multocida, Haemophilusbacteria, for example Haemophilus influenzae, and Bordetella bacteria,for example B. bronchiseptica (B.=Bordetella) Bacteriodacea, such asBacteroides bacteria, for example Bacteroides fragilis, Fusiformebacteria for example Fusobacterium fusiforme and Sphaerophorus bacteriafor example Sphaerophorus necrophorus, Sph. necroticus and Sph.pyrogenes (Sph.=Sphaerophorus); Bacillaceae, such as aerobicspore-forming Bacillaceae for example Bacillus anthracis, B. subtilisand B. cereus (B.=Bacillus), and anaerobic spore-forming Clostridia, forexample clostridium perfringens.

The above list of pathogens is purely by way of example and is in no wayto be interpreted as restrictive.

Examples which may be mentioned of diseases which can be prevented,alleviated and/or cured by the active compounds according to theinvention are: diseases of the respiratory passages and of thepharyngeal cavity; otitis; pharyngitis; pneumonia; peritonitis;pyelonephritis; cystitis; endocarditis; systemic infections; bronchitisand arthritis.

As stated above, the invention also relates to the use in human andveterinary medicine of the compounds of the invention.

The present invention provides a pharmaceutical composition containingas active ingredient a compound of the invention in admixture with asolid or liquefied gaseous diluent, or in admixture with a liquiddiluent other than a solvent of a molecular weight less than 200(preferably less than 350) except in the presence of a surface activeagent.

The invention further provides a pharmaceutical composition containingas active ingredient a compound of the invention in the form of asterile and/or isotonic aqueous solution.

The invention also provides a medicament in dosage unit form comprisinga compound of the invention.

The invention also provides a medicament in the form of tablets(including lozenges and granules), dragees, capsules, pills, ampules orsuppositories comprising a compound of the invention.

"Medicament" as used in this specification means physically discretecoherent portions suitable for medical administration. "Medicament indosage unit form" as used in this specification means physicallydiscrete coherent units suitable for medical administration eachcontaining a daily dose or a multiple (up to four times) or sub-multiple(down to a fortieth) of a daily dose of the compound of the invention inassociation with a carrier and/or enclosed within an envelope. Whetherthe medicament contains a daily dose or, for example, a half, a third,or a quarter of a daily dose will depend on whether the medicament is tobe administered once or, for example, twice, three times or four times aday respectively.

The pharmaceutical compositions according to the invention may, forexample, take the form of ointments, gels, pastes, creams, sprays(including aerosols), lotions, suspensions, solutions and emulsions ofthe active ingredient in aqueous or non-aqueous diluents, syrups,granulates or powders.

The diluents to be used in pharmaceutical compositions (e.g. granulates)adapted to be formed into tablets, dragees, capsules and pills includethe following: (a) fillers and extenders, e.g. starch, sugars, mannitol,and silicic acid; (b) binding agents, e.g. carboxymethyl cellulose andother cellulose derivatives, alginates, gelatine and polyvinylpyrrolidone; (c) moisturizing agents e.g. glycerol; (d) disintegratingagents, e.g. agar-agar calcium carbonate and sodium bicarbonate; (e)agents for retarding dissolution e.g. paraffin; (f) resorptionaccelerators, e.g. quaternary ammonium compounds; (g) surface activeagents, e.g. cetyl alcohol, glycerol monostearate; (h) adsorptivecarriers, e.g. kaolin and bentonite; (i) lubricants, e.g. talc, calciumand magnesium stearate and solid polyethylene glycols.

The tablets, dragees, capsules and pills formed from the pharmaceuticalcompositions of the invention can have the customary coatings, envelopesand protective matrices, which may contain opacifiers. They can be soconstituted that they release the active ingredient only or preferablyin a particular part of the intestinal tract, possibly over a period oftime. The coatings, envelopes and protective matrices may be made, forexample, of polymeric substances or waxes.

The ingredient can also be made up in microencapsulated form togetherwith one or several of the above mentioned diluents.

The diluents to be used in pharmaceutical compositions adapted to beformed into suppositories can, for example, be the usual water-solubleor water-insoluble diluents, such as polyethylene glycols and fats (e.g.cocoa oil and high esters [e.g. C₁₄ -alcohol with C₁₆ -fatty acid]) ormixtures of these diluents.

The pharmaceutical compositions which are ointments, pastes, creams andgels can, for example, contain the usual diluents, e.g. animal andvegetable fats, waxes, paraffins, starch, tragacanth, cellulosederivatives, polyethylene glycols, silicones, bentonites, silicic acid,talc and zinc oxide or mixtures of these substances.

The pharmaceutical compositions which are powders and sprays can, forexample, contain the usual diluents, e.g. lactose, talc, silicic acid,aluminum hydroxide, calcium silicate, and polyamide powder or mixturesof these substances. Aerosol sprays can, for example, contain the usualpropellants, e.g. chlorofluorohydrocarbons.

The pharmaceutical compositions which are solutions and emulsions can,for example, contain the customary diluents (with, of course, the abovementioned exclusion of solvents having a molecular weight below 200except in the presence of a surface-active agent), such as solvents,dissolving agents and emulsifiers; specific examples of such diluentsare water, ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethylacetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butyleneglycol, dimethylformamide, oils [for example ground nut oil], glycerol,tetrahydrofurfuryl alcohol, polyethylene glycols and fatty acid estersof sorbitol or mixtures thereof.

For parenteral administration, solutions and emulsions should besterile, and, if appropriate, blood-isotonic.

The pharmaceutical compositions which are suspensions can contain theusual diluents, such as liquid diluents, e.g. water, ethyl alcohol,propylene glycol, surface-active agents (e.g. ethoxylated isostearylalcohols, polyoxyethylene sorbite and sorbitan esters), microcrystallinecellulose, aluminium metahydroxide, bentonite, agar-agar and tragacanthor mixtures thereof.

All the pharmaceutical compositions according to the invention can alsocontain coloring agents and preservatives as well as perfumes andflavoring additions (e.g. peppermint oil and eucalyptus oil) andsweetening agents (e.g. saccharin).

The pharmaceutical compositions according to the invention generallycontain from 0.1 to 99.5, usually from 0.5 to 95% of the activeingredient by weight of the total composition.

In addition to a compound of the invention, the pharmaceuticalcompositions and medicaments according to the invention can also containother pharmaceutically active compounds. They may also contain aplurality of compounds of the invention.

Any diluent in the medicaments of the present invention may be any ofthose mentioned above in relation to the pharmaceutical compositions ofthe present invention. Such medicaments may include solvents ofmolecular weight less than 200 as sole diluent.

The discrete coherent portions constituting the medicament according tothe invention will generally be adapted, by virtue of their shape orpackaging, for medical administration and may be, for example, any ofthe following: tablets, (including lozenges and granulates), pills,dragees, capsules, suppositories and ampules. Some of these forms may bemade up for delayed release of the active ingredient. Some, such ascapsules, include a protective envelope which renders the portions ofthe medicament physically discrete and coherent.

The preferred daily dose for administration of the medicaments of theinvention is from 300 mg to 40 g, preferably from 750 mg to 15 g, ofactive ingredient.

The production of the above mentioned pharmaceutical compositions andmedicaments is carried out by any method known in the art, for example,by mixing the active ingredient(s) with the diluent(s) to form apharmaceutical composition (e.g. a granulate) and then forming thecomposition into the medicament (e.g. tablets).

This invention further provides a method of combating (includingprevention, relief and cure of) the above mentioned diseases in humanand non-human animals, which comprises administering to the animals acompound of the invention alone or in admixture with a diluent or in theform of a medicament according to the invention.

It is envisaged that these active compounds will be administeredperorally, parenterally (for example intramuscularly, intraperitoneallyor intravenously), rectally or locally, preferably parenterallyespecially intravenously or intramuscularly. Preferred pharmaceuticalcompositions and medicaments are therefore those adapted for parenteraladministration, such as injection solutions and suspensions.Administration in the method of the invention is preferably parenteral.

In general it has proved advantageous to administer amounts of from 6 mgto 800 mg, preferably from 15 to 300 mg, kg of body weight per day toachieve effective results. Nevertheless, it can at times be necessary todeviate from those dosage rates, and in particular to do so as afunction of the nature and body weight of the human or animal subject tobe treated, the individual reaction of this subject to the treatment,the type of formulation in which the active ingredient is administeredand the mode in which the administration is carried out and the point inthe progress of the disease or interval at which it is to beadministered. Thus it may in some case suffice to use less than theabove mentioned minimum dosage rate, while in other cases the upperlimit mentioned must be exceeded to achieve the desired results. Wherelarger amounts are administered it can be advisable to divide these intoseveral, for example 3, individual administrations over the course ofthe day. Advantageously each individual administration contains from 2to 300, preferably from 10 to 150, mg/kg of body weight.

When used as feedstuff additives, the new compounds can be administeredin the customary manner together with the feedstuff or with feedstuffformulations or with the drinking water. By this means it is possible toprevent an infection by Gram-negative or Gram-positive bacteria and alsoto achieve better utilization of the feedstuff.

The new β-lactam compounds are distinguished by powerful antibacterialactions, which have been tested in vivo and in vitro, and by oralresorbability.

The β-lactam compounds according to the invention can, in order tobroaden the spectrum of action or to achieve a more powerful action,also be combined with, for example, amino-glycoside antibiotics, such asgentamycin, sisomycin, kanamycin, amicacin or tobramycin.

Isoxazolyl-penicillins (for example oxacillin and dicloxacillin) canalso be used as combination partners.

The activity of the β-lactam compounds according to the invention can bedemonstrated, by way of example, by the following in vitro and in vivoexperiments:

(a) In vitro Experiment

The compounds from Examples 2, 5 and 6 hereinbelow which can be regardedas typical representatives of the compounds according to the invention,were diluted to a content of 100 μg/ml with Muller-Hinton nutrientbroth. In each case, the nutrient solution contained 1×10⁵ to 2×10⁵bacteria per milliliter. The small tubes containing this batch were ineach case incubated for 24 hours and the degree of turbidity was thendetermined. Freedom from turbidity indicates action. At a dosage of 100μg/ml, the following bacterial cultures were free from turbidity(sp.=species):

Klebsiella pneumoniae; Enterobacter aerogenes sp.; Serratia marcescens;Escherichia coli BE; Salmonella sp.; Shigella sp.; Proteus,indole-negative and indole-positive sp.; Pasteurella pseudotuberculosis;Haemophilus influenzae; Bordetella bronchiseptica; Staphylococcus aureus133; Neisseria catarrhalis sp.; Diplococcus pneumoniae sp.;Streptococcus pyogenes, W.; Enterococcus sp.; Lactobacillus sp.;Corynebacterium diphtheriae gravis; Cornyebacterium pyogenes M;Clostridium botulinum and Clostridium tetani.

(b) In vivo Experiment

Table 1 which follows shows the action of one of the new β-lactamcompounds, which can be regarded as typical compounds according to theinvention, against some bacteria listed in an animal experiment usingwhite mice. White mice of the CF₁ strain were infected intraperitoneallywith the particular strain of bacteria indicated.

                  TABLE 1                                                         ______________________________________                                         Animal experiment using white mice                                           ______________________________________                                        Determination of the ED.sub.100 after 24 hours                                                Dose in mg of the compound from                                               Example 2 per kg/body weight                                  Germ            (subcutaneously)                                              ______________________________________                                        Escherichia coli C 165                                                                        1 × 200                                                 Klebsiella      2 × 150                                                 ______________________________________                                        Therapy:                                                                             1 administration:                                                                          30 minutes after infection                                       2 administrations:                                                                         (a) 30 minutes after infection                                                (b) 90 minutes after infection                            ______________________________________                                    

The ED₁₀₀ is the dose at which 100% of the infected animals stillsurvive after 24 hours.

The process according to the invention may be illustrated by theexamples which follow:

The α-aminobenzylpenicillin used in the examples which follow containsabout 14% of water, but anhydrous α-aminobenzylpenicillin (compare U.S.Pat. No. 3,144,445) can also equally well be used.

The 7-(α-amino-phenylacetamido)-3-acetoxymethyl-ceph-3-em-4-carboxylicacid used in the examples contains 8% of water, but anhydrous7-(α-amino-phenylacetamido)-3-acetoxymethyl-ceph-3-em-4-carboxylic acidcan also equally well be used.

The water content of the starting compounds is irrelevant in carryingout the process according to the invention.

By "ampicillin" is meant that α-aminobenzylpenicillin with theD═R-configuration in the side chain.

By "cephaloglycine" is meant that7-(α-amino-phenylacetamido)-3-acetoxymethyl-ceph-3-em-4-carboxylic acidwith the D═R-configuration in the side chain.

The symbols in the NMR (Nuclear Magnetic Resonance) spectrum date quotedhereinbelow have the following meanings:

s=singlet

AB=AB system

sept.=septet

Explanation of the abbreviations used in the examples:

pts. by wt.=parts by weight

pts. by vol.=parts by volume

hrs.=hours

hr.=hour

THF=tetrahydrofuran

ethyl acetate=acetic acid ethyl ester

room temperature=about 20° C.

abs.=absolute

decomp. pt.=decomposition point

The percentage data for the yields denote yields in percent of theory.

EXAMPLE 1 (Intermediate) ##STR48##

First 6.3 pts. by wt. of phosgene and then 11.8 pts. by wt. oftri-n-butylamine are added to a suspension of 8.0 pts. by wt. of1-propenyl-2-oxo-imidazolidine (prepared according to DAS (GermanPublished Specification) No. 1,057,126) in 100 pts. by vol. of abs.ethyl acetate. The reaction mixture is stirred overnight andconcentrated. The 1-chlorocarbonyl-2-oxo-3-propenyl-imidazolidine formedis used for Example 2 in a mixture with tributylammonium chloride. IR(liquid paraffin): 1,800 and 1,710 cm⁻¹.

EXAMPLE 2 ##STR49##

25.6 pts. by wt. of ampicillin trihydrate are dissolved in 150 pts. byvol. of 80 percent strength aqueous THF, at 5°-10°, by adding 4 percentstrength sodium hydroxide solution. During this procedure, the pH valueshould not exceed 8.3. 31.4 pts. by wt. of1-chlorocarbonyl-2-oxo-3-propenyl-imidazolidine (1:1 mixture withtributylammonium chloride, from Example 1), dissolved in 100 pts. byvol. of THF, are added dropwise to this solution, the pH being keptbetween 7.3 and 7.5 by successive addition of 2 percent strength sodiumhydroxide solution. When no further sodium hydroxide solution isconsumed, 250 pts. by vol. of water are added and the THF is strippedoff. The aqueous phase is extracted with ethyl acetate, separated off,cooled to 5°, covered with a layer of cold ethyl acetate and acidified.The aqueous phase is extracted by shaking several times with ethylacetate and the combined ethyl acetate phases are dried over magnesiumsulphate and concentrated. The residue is dissolved in 100 pts. by vol.of cold methanol, 49 pts. by vol. of a 1 M methanolic sodium caprylatesolution are added and the mixture is concentrated to half the volume.Sodium{D-α[(2-oxo-3-propenyl-imidazolidin-1-yl)-carbonylamino]phenylacetamido}-penicillanateis precipitated by adding ether which contains 5% of methanol. 20.0 pts.by wt. of decomp. pt. 170°-175° are obtained. IR (KBr): 1,770, 1,715,1,660, 1,605, 1,520, 1,400 and 1,255 cm⁻¹.

The product is present partially in the hydrated form.

EXAMPLE 3 (INTERMEDIATE) ##STR50##

1-Isobutenyl-2-oxo-imidazolidine is prepared analogously to1-propenyl-2-oxo-imidazolidine. Boiling point ₀.4mm 152° melting point86°-90°.

IR (CHCl₃): 3,260, 1,695, 1,485, 1,425, 1,280 and 1,205 cm⁻¹.

NMR (CDCl₃): s, broad 6.43 (NH), sept. 5.94 (1H), A₂ B₂ centred at 3.67(4H), s 1.72 (3H), and s 1.69(3H) ppm (ε).

EXAMPLE 4 (INTERMEDIATE) ##STR51##

5.0 pts. by wt. of 1-isobutenyl-2-oxo-imidazolidine, 3.5 pts. by wt. ofphosgene and 6.6 pts. by wt. of tri-n-butylamine are reacted as inExample 1. The 1-chlorocarbonyl-2-oxo-isobutenyl-imidazolidine formed isused for Example 5, together with tributylammonium chloride.

IR (liquid paraffin): 1,800 and 1,720 cm⁻¹.

EXAMPLE 5 ##STR52##

36.0 pts. by wt. of ampicillin trihydrate in 300 pts. by vol. of 80percent strength aqueous THF are reacted with 18.1 pts. by wt. of a 1:1mixture of 1-chlorocarbonyl-2-oxo-3-isobutenyl-imidazolidine andtributylammonium chloride as in Example 2. 11.6 pts. by wt. of sodium6-{D-α[(2-oxo-3-isobutenyl-imidazolin-1-yl)-carbonylamino]-phenylacetamido}penicillanateof decomp. pt. 180°-185° are obtained.

IR (KBr): 1,770, 1,720, 1,665, 1,605, 1,530 and 1,250 cm⁻¹.

The product is present mostly in the hydrated form: ##STR53##

EXAMPLE 6

The following compound was prepared in the same manner as Example 5,starting from cephaloglycine dihydrate: ##STR54##

Analogously to Example 5, the compound is present mostly in the hydratedform.

It will be appreciated that the instant specification and examples areset forth by way of illustration and not limitation, and that variousmodifications and changes may be made without departing from the spiritand scope of the present invention.

What is claimed is:
 1. A compound of the formula ##STR55## in which B isphenyl, hydroxyphenyl or 1,4-cyclohexadien-1-yl,Z is vinyl, propenyl,isopropenyl, isobutenyl, sec.-butenyl, or 1-methylisobutenyl, or ahydrated form thereof,or a pharmaceutically acceptable salt thereof. 2.A compound according to claim 1 of the formula ##STR56## or a saltthereof.
 3. A compound according to claim 1 of the formula ##STR57## ora salt thereof.
 4. A compound according to claim 1 of the formula##STR58## or a salt thereof
 5. A compound according to claim 1 of theformula ##STR59## or a salt thereof.
 6. A pharmaceutical compositioncontaining as an active ingredient an antibacterially effective amountof a compound or salt thereof according to claim 1 in admixture with apharmaceutically acceptable diluent.
 7. A medicament in dosage unit formcomprising a composition according to claim
 6. 8. A method of combatingbacterial diseases in human and non-human animals which comprisesadministering to the animals an antibacterially effective amount of acompound or salt thereof according to claim
 1. 9. A method of promotinggrowth and improving feedstuff utilization in non-human animals whichcomprises administering to the animals a growth promoting amount of anactive compound or salt thereof according to claim
 1. 10. An animalfeedstuff comprising feed and a compound or a salt thereof according toclaim
 1. 11. A method of preserving inorganic or organic materials whichcomprises applying to the materials a preservation-effective amount of acompound or salt thereof according to claim 1.